本文選題：三聚氰胺　切入點(diǎn)：三聚氰酸　出處：《沈陽(yáng)農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：三聚氰胺(Melamine)是典型的三嗪類有機(jī)雜環(huán)化合物,它作為一種化工原料,被廣泛應(yīng)用于塑料、化肥等工業(yè)生產(chǎn)。三聚氰酸(Cyanuric acid)是三聚氰胺經(jīng)連續(xù)脫氨基反應(yīng)生成的水解產(chǎn)物。目前,國(guó)內(nèi)外對(duì)于該類物質(zhì)的研究主要集中在對(duì)機(jī)體器官、組織所造成的腎毒性、肝毒性、生殖毒性和遺傳毒性等方面,關(guān)于它們對(duì)機(jī)體免疫系統(tǒng)的毒性作用研究還較為有限,能夠調(diào)節(jié)該類物質(zhì)毒性的天然解毒劑的研究是目前研究熱點(diǎn)。本論文旨在研究三聚氰胺及其與三聚氰酸聯(lián)合對(duì)雄性小鼠免疫功能的影響,同時(shí)探索了茶多酚、Vitamin C和Vitamin E作為天然解毒劑對(duì)該類物質(zhì)所起的調(diào)節(jié)作用。試驗(yàn)一茶多酚、Vitamin C和Vitamin E對(duì)三聚氰胺及其與三聚氰酸聯(lián)合攻毒小鼠B細(xì)胞的調(diào)節(jié)作用研究方法:將56只雄性昆明小鼠隨機(jī)分為三聚氰胺組(30mg.kg-1)、三聚氰胺+茶多酚組(30+400mg.kg-1)、三聚氰胺+Vitamin C 和 Vitamin E 組(30+215/286mg.kg-1)、三聚氰胺+三聚氰酸聯(lián)合組(15+15mg.kg-1)、聯(lián)合+茶多酚組(15+15+400mg.kg-1)、聯(lián)合+Vitamin C和Vitamin E組(15+15+215/286mg.kg-1)及空白對(duì)照組7個(gè)試驗(yàn)組。小鼠隔天灌胃28d,腹腔注射1%綿羊紅細(xì)胞用于激發(fā)機(jī)體免疫反應(yīng)。眼球采血和頸椎脫臼處死小鼠,采用全自動(dòng)血液分析儀和流式細(xì)胞儀檢測(cè)小鼠外周血和漿細(xì)胞表面標(biāo)志CD138+的表達(dá)率。無(wú)菌摘取脾臟,RT-PCR、Western Blot法檢測(cè)Blimp-1基因和蛋白的相對(duì)表達(dá)量。結(jié)果表明:①與對(duì)照組相比,聯(lián)合組、聯(lián)合+茶多酚及聯(lián)合+VitaminC和VitaminE組的白細(xì)胞的數(shù)量均顯著升高(P0.05),而聯(lián)合+茶多酚組小鼠外周血中白細(xì)胞的數(shù)量顯著低于聯(lián)合組(P0.05)。②從小鼠淋巴細(xì)胞的數(shù)量上看,聯(lián)合組和聯(lián)合+VitaminC和Vitamin E組與對(duì)照組比數(shù)值顯著增加(P0.05),而聯(lián)合+茶多酚組未出現(xiàn)顯著性差異(P0.05)。③三聚氰胺組和聯(lián)合組CD138+的表達(dá)率減少(P0.01),而其它試驗(yàn)組未呈現(xiàn)出顯著性變化(P0.05)。④從小鼠脾臟內(nèi)Blimp-1蛋白的相對(duì)表達(dá)量來(lái)看,聯(lián)合攻毒組與空白對(duì)照組小鼠相比極顯著下降(P0.01),而其它試驗(yàn)組無(wú)顯著性差異(P0.05);各試驗(yàn)組與空白對(duì)照組比,Blimp-1 mRNA相對(duì)表達(dá)量并未表現(xiàn)出顯著性的變化(P0.05)。說(shuō)明該類物質(zhì)能通過(guò)降低Blimp-1蛋白的表達(dá)量,使B淋巴細(xì)胞的成熟和分化受阻,而茶多酚、Vitamin C和Vitamin E可以在一定程度上調(diào)節(jié)這種抑制作用。試驗(yàn)二茶多酸、Vitamin C和Vitamin E對(duì)三聚氰胺及其與三聚氰酸聯(lián)合攻毒小鼠T細(xì)胞的調(diào)節(jié)作用研究方法:試驗(yàn)動(dòng)物分組及處理同試驗(yàn)一,流式細(xì)胞儀檢測(cè)小鼠外周血中T細(xì)胞表面標(biāo)志 CD3+、CD4+、CD8+,及小鼠 CTL 細(xì)胞表面標(biāo)志 CD8a+CD44hiCD62Llow。RT-PCR、Western Blot檢測(cè)Notch-1基因和蛋白的相對(duì)表達(dá)情況。結(jié)果表明:①聯(lián)合攻毒組小鼠CD4+/CD8+的比值與空白對(duì)照組相比顯著降低(P0.05),而聯(lián)合+茶多酚組、聯(lián)合+Vitamin C和Vitamin E組小鼠CD4+/CD8+的比值變化不顯著(P0.05),CD4+與CD8+的比值降低意味著機(jī)體的免疫功能可能在一定程度上受到了抑制,而使用了茶多酚、VitaminC和VitaminE后比值得到恢復(fù)。②各試驗(yàn)組與空白對(duì)照組相比較小鼠外周血中的CTL細(xì)胞表達(dá)率并未出現(xiàn)顯著性差異(P0.05),但均低于對(duì)照組。③聯(lián)合攻毒組Notch-1基因的相對(duì)表達(dá)量較空白對(duì)照組極顯著升高(P0.01),而聯(lián)合+茶多酚組與聯(lián)合組相比Notch-1 mRNA的相對(duì)表達(dá)量顯著降低(P0.05),說(shuō)明該類物質(zhì)通過(guò)使Notch-1基因相對(duì)表達(dá)量的增多來(lái)調(diào)控機(jī)體內(nèi)T細(xì)胞的分化和成熟。但三聚氰胺或者其與三聚氰酸聯(lián)合作用于機(jī)體時(shí),VitaminC和VitaminE會(huì)使Notch-1蛋白相對(duì)表達(dá)量顯著升高(P0.05),其它試驗(yàn)組并沒(méi)有發(fā)生顯著變化(P0.05),這與mRNA的檢測(cè)結(jié)果不一致,其原因還有待于進(jìn)一步研究。以上研究說(shuō)明茶多酚、Vitamin C和Vitamin E對(duì)該類物質(zhì)的免疫毒性具有一定的調(diào)節(jié)作用。
[Abstract]:Melamine (Melamine) is a typical three triazine heterocyclic organic compounds, it is used as a chemical raw material, widely used in plastics, fertilizers and other industrial production. Cyanuric acid (Cyanuric acid) is the melamine continuous de hydrolysate amino reaction. At present, domestic and foreign research on this kind of material is mainly concentrated on the body organs, tissue caused by kidney toxicity, liver toxicity, reproductive toxicity and genetic toxicity, about them on immune toxicity study is still limited, can adjust the toxic substance natural antidote research is the current research focus. This paper aims to study the melamine and cyanuric acid and its influence on the immune function of male mice, and explore the role of tea polyphenols, Vitamin C and Vitamin E as a natural antidote to this kind of material. The test of tea polyphenols, Vit Effect of Amin C and Vitamin E methods of melamine and cyanuric acid combined with cytotoxic attack B mice: 56 male Kunming mice were randomly divided into group (30mg.kg-1), melamine, melamine + tea polyphenol group (30+400mg.kg-1), +Vitamin C and Vitamin E group of melamine (30+215/286mg.kg-1), melamine + cyanuric acid group (15+15mg.kg-1 +), combined with tea polyphenols (15+15+400mg.kg-1), C and Vitamin combined with +Vitamin E group (15+15+215/286mg.kg-1) and blank control group 7 experimental groups. The mice were gavaged with 28d, abdominal cavity injection of 1% sheep red blood cells to stimulate immune response. The eye blood and the mice were killed by cervical dislocation by automatic blood analyzer and flow cytometry in peripheral blood and plasma cell surface marker CD138+ expression rate. Sterile removal of spleen, RT-PCR, detection of Blimp-1 Western by Blot gene and protein The relative expression level. The results showed that: compared with the control group, the combined group, the number of white blood cells + combined with tea polyphenols and the combination of +VitaminC and VitaminE group were significantly increased (P0.05), and combined with tea polyphenols + white cells in peripheral blood of mice was significantly lower than the number of combined group (P0.05) from the. The number of lymphocytes of mice at the combined group and the combination of +VitaminC and Vitamin in E group and control group increased significantly (P0.05), and combined with numerical + TP group did not show significant difference (P0.05). The expression of melamine group and combined group CD138+ (P0.01), and reduce the rate of other groups did not show up significant changes (P0.05). The Blimp-1 from the spleen of mice in the expression of the protein, combined with poison attack group and blank control group mice decreased significantly (P0.01), while there was no significant difference between the other experimental group (P0.05); the experimental group compared with blank control group, Blimp-1 M The relative expression of RNA did not show significant changes (P0.05). The material can reduce the expression of Blimp-1, B lymphocyte maturation and differentiation was blocked, and tea polyphenols, Vitamin C and Vitamin E can inhibit the regulation in a certain extent. Test two theapolyacid, regulation to study the effect of Vitamin C and Vitamin E methods of melamine and cyanuric acid combined with cytotoxic attack T mice: experimental animal groups and treatments were same, flow cytometry in mice T cell surface markers in peripheral blood CD3+, CD4+, CD8+, and mouse CTL cell surface marker CD8a+CD44hiCD62Llow.RT-PCR, the relative expression of Western Blot detection of Notch-1 gene and protein. The results showed that the combined ratio and blank mice challenged group CD4+/CD8+ compared with the control group decreased significantly (P0.05), and combined with tea polyphenols + group, combined with +Vitamin C and V The change of the ratio of itamin in E group CD4+/CD8+ was not significant (P0.05), the ratio of CD4+ and CD8+ mean lower immune function may to some extent inhibited, and the use of tea polyphenols, recovery ratio of VitaminC and VitaminE. The expression of the test group and blank control group compared with mouse peripheral CTL in the blood cell rate did not show significant difference (P0.05), but were lower than the control group. The combined inoculation group the expression of Notch-1 was significantly higher than the control group (P0.01), and the joint + TP group and combination group compared the relative expression of Notch-1 mRNA decreased significantly (P0.05) this kind of material that, by making the Notch-1 relative expression increased to differentiation and maturation of T cells in the body. But the regulation of melamine and cyanuric acid and its effect on the body, VitaminC and VitaminE can make Notch-1 protein. The expression increased significantly (P0.05), the other experimental group did not change significantly (P0.05), this inconsistency detection and mRNA results, the reason remains to be further studied. The above research shows that tea polyphenols, Vitamin C and Vitamin E has a certain effect on regulating the immune toxicity of the substances.

【學(xué)位授予單位】：沈陽(yáng)農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S859.8
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本文編號(hào)：1664691